Identification of fentanyl analogs and potential biomarkers in urine using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and liquid chromatography-quadrupole/time of flight mass spectrometry (LC-Q/TOF-MS)

Novel synthetic opioids are a class of drugs abused for their potent analgesic effect and are responsible for many fatal intoxications, particularly within the United States. A targeted assay was developed and validated using LC-MS/MS, capable of identifying nineteen fentalogs. Solid phase extraction was used to isolate analytes of interest from urine. Limits of detection ranged from 0.05 to 0.1 ng/mL and the limit of quantitation was 0.5 ng/mL. Extraction efficiencies using the optimized procedure were 77-88 % for all targeted species. Bias, precision, matrix effects and interferences were within acceptable thresholds for all analytes. The validated assay was used to identify analytes of interest from thirty-seven individuals that had used fentanyl and related substances. In addition to quantitative analyses, a non-targeted liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q/TOF-MS) assay was also used to identify additional substances and potential biomarkers. Additional N-oxide and N-dealkylated species were identified using this approach, and the potential for biomarker use is presented, given the stability of some analytes within this class.